Abstract
When subjected to hyperosmotic pressure resulting from NaCl addition, KR12H-2 transfectoma, like most hybridomas, displayed a decrease in specific growth rate (mu) and an increase in specific antibody productivity (q(Ab)). Elevation of medium osmolality from 285 to 425 mOsm/kg decreased mu by 20%, while it increased q(Ab) by 376%. Although cell mass also increased at higher osmolality, it was not the main factor in increasing q(Ab). Hyperosmotic pressure was found to enhance transcription levels of immunoglobulin (Ig) mRNAs preferentially, compared with non-IgG mRNA. The transcription levels of both heavy chain (HC) and light chain (LC) mRNAs were enhanced as much as q(Ab). This result suggests that enhanced q(Ab) at higher osmolality was mainly due to enhanced transcription levels of Ig mRNA. However, these increased transcription levels of Ig mRNAs were not due to the enhanced stability of Ig mRNA. In fact, the stability of Ig mRNAs decreased at higher osmolality. Elevation of osmolality from 285 mOsm/kg to 425 mOsm/kg decreased the half-lives of HC and LC mRNAs by 37% and 36%, respectively. A simple mathematical model revealed that transcription rates of Ig mRNAs increased by more than 476% at 425 mOsm/kg. These elevated transcription levels could, in turn, increase the translation rates of Ig polypeptides. However, the translation rates of Ig polypeptides were not enhanced as much as the transcription levels of Ig mRNAs and q(Ab). The elevation of osmolality from 285 mOsm/kg to 425 mOsm/kg increased HC and LC mRNA specific translation rates by 172% and 240%, respectively. Taken together, the data suggest that (1) enhanced q(Ab) of KR12H-2 transfectoma at higher osmolality is due to elevated transcription rates of Ig mRNAs and expedited post-translational processing of Ig, and (2) antibody secretion by KR12H-2 transfectoma is most likely controlled at the level of Ig translation, particularly HC translation.
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